Alizadeh-Khiavi K, Ali-Khan Z
Department of Microbiology and Immunology, McGill University, Montreal, Quebec, Canada.
Br J Exp Pathol. 1988 Oct;69(5):605-19.
Low ionic strength acidic buffer, Sephadex G-200 and Benzamidine-Sepharose (BZ) gel chromatography, have been used for the partial purification of alveolar hydatid cyst (AHC) induced amyloid enhancing factor (AEF). BZ-gel bound AEF (AEF-BZ) demonstrated AEF activity in the mouse bioassay, proteolytic activity against Hide powder azure showed two major and three minor peptides on SDS-PAGE. Pretreatment of AEF-BZ with 10 mM phenylmethylsulphonyl fluoride or 20 mM p-chloromercuribenzoic acid completely abolished its bioactivity in vivo and proteolytic activity in vitro. Polyclonal anti-AEF antibody (AAA) was generated which on passive transfer into mice completely abolished the bioactivity of both casein-induced, or AHC-induced AEF. The AAA absorbed on Sepharose gel conjugated to normal mouse serum developed one common precipitin band between AE and AEF-positive sera from AHC-infected and old retired mice and in immunostaining it bound to the cytoplasmic granular components of a majority of splenic and peritoneal leucocytes from AHC-infected mice. In contrast, only a few normal mouse leucocytes showed positive staining. We suggest that AEF, in all probability, is a serine/thiol protease of leucocyte origin whose intracellular and humoral concentrations increase significantly during amyloidosis. The role of lysosomal proteases and anti-AEF antibody which has been successfully generated for the first time is discussed with reference to the origin of AEF and its presumed biological function in amyloidogenesis.
低离子强度酸性缓冲液、葡聚糖凝胶G - 200和苯甲脒 - 琼脂糖凝胶（BZ）柱色谱法已用于肺泡包虫囊肿（AHC）诱导的淀粉样蛋白增强因子（AEF）的部分纯化。结合在BZ凝胶上的AEF（AEF - BZ）在小鼠生物测定中表现出AEF活性，对皮粉天青有蛋白水解活性，在SDS - PAGE上显示出两条主要肽段和三条次要肽段。用10 mM苯甲基磺酰氟或20 mM对氯汞苯甲酸预处理AEF - BZ可完全消除其体内生物活性和体外蛋白水解活性。制备了多克隆抗AEF抗体（AAA），将其被动转移到小鼠体内可完全消除酪蛋白诱导的或AHC诱导的AEF的生物活性。吸附在与正常小鼠血清偶联的琼脂糖凝胶上的AAA在AHC感染的和老龄退休小鼠的AE和AEF阳性血清之间产生一条共同沉淀带，在免疫染色中它与AHC感染小鼠的大多数脾和腹膜白细胞的细胞质颗粒成分结合。相比之下，只有少数正常小鼠白细胞呈阳性染色。我们认为，AEF很可能是一种源自白细胞的丝氨酸/硫醇蛋白酶，其细胞内和体液浓度在淀粉样变性期间显著增加。参照AEF的起源及其在淀粉样蛋白形成中假定的生物学功能，讨论了首次成功产生的溶酶体蛋白酶和抗AEF抗体的作用。
